Extended Data Fig. 5: Additional data for Drosophila tau RNAi studies.
From: Tau is required for glial lipid droplet formation and resistance to neuronal oxidative stress
(a) UAS-Control (Ctrl) RNAi or UAS-tau RNAi was expressed ubiquitously using da-GAL4. Protein extracted from 2d heads was analyzed by WB for dTau levels. A datapoint represents the relative band density per technical replicate (n). n = 6. (b) UAS-Control (LacZ), UAS-Ctrl RNAi or UAS-tau RNAi were expressed in adult flies starting at 2d using the pan-neuronal, drug inducible driver, elavGS. n = 234-325 per genotype. (c) UAS-Ctrl RNAi or UAS-tau RNAi were expressed through development and adulthood using the pan-neuronal driver, elav-GAL4, at 29 °C. Control RNAi n = 161, tau RNAi n = 125. As the GAL4/UAS system is temperature-dependent, this produces higher RNAi expression than standard 25 °C rearing temperatures (see Fig. 3e). (b-c) Survival was measured using Kaplan-Meier plots. P-value ≤ 0.0001 seen with UAS-tau RNAi versus UAS- Ctrl RNAi in b and c, with UAS-tau RNAi versus UAS-Control in c, and UAS- Ctrl RNAi versus UAS-Control in c. UAS-Ctrl RNAi and UAS-tau RNAi are in a w+ background. UAS-Control is in a w* background. (d) UAS-Control (GFP) RNAi-2 or UAS-tau RNAi-2 were expressed in retinal glia using 54C-GAL4. Rh:ND42 RNAi was used to induce neuronal ROS and glial LDs. LDs were visualized in fly retinas from 2d animals using BODIPY 493/503 (white puncta, arrows). A datapoint represents the average LD number per ommatidium per animal (n). n = 6. (e) UAS-Control RNAi or UAS-tau RNAi was expressed in retinal glia using 54C-GAL4 and TEM was performed to visualize morphological changes in the fly retina. Glial cells = green. Shown are representative images for the quantification shown in Fig. 5d. (f) UAS-Control RNAi or UAS-tau RNAi were expressed in glia of 1-2d adult flies for 10d. Fixed brains were stained for LDs using BODIPY 493/503 (white puncta, arrows). Shown are representative images for the quantification shown in Fig. 5f. (g) The total levels of triglycerides (TG) were measured in whole heads from either 10d or 26d flies expressing UAS-RNAi in adult glia using repoGS. A datapoint represents the relative TG levels per technical replicate (n). n = 8. Statistics: unpaired 2-sided student t-test (a, d), Log-rank test (b, c), 1-way ANOVA with Tukey’s multiple comparisons test (g). P-values = not significant (n.s.) >0.05, *≤0.05, **≤0.01, ***≤0.001, ****≤0.0001. Data presented as mean ± SEM. See Supporting Data 1 for details on technical replicates, n, and P-values.
